黄瓜霜霉病菌侵染若干因子的研究

研究了温、湿度条件对黄瓜霜霉病菌致病性的影响,结果表明,25～35℃最适宜黄瓜霜霉病的发生,15/35℃的交替温度变化最有利于霜霉病菌的侵染,但35℃以上的高温对霜霉病菌具有杀伤作用;2 h的湿度条件就足以引起侵染,一旦侵入寄主,环境湿度条件对病害的发展影响不大.-20℃低温冷冻保存10个月和干燥放置7 d的霜霉菌种仍具致病力.发病的黄瓜叶片可以连续产生孢子囊,但随着发病时间的延长产生孢子囊量逐渐减少.活体叶片单位面积上产生的孢子囊量比离体叶片大,且显症天数与叶片单位面积产生孢囊量呈抛物线型关系.     

大白菜霜霉病菌寄生霜霉孢子囊的保藏方法

通过离体子叶冷冻法、离体叶片冷冻法和10%二甲基亚砜+5%脱脂乳冷冻法等3种方法来保藏大白菜霜霉病菌寄生霜霉,并采用离体子叶接种法测定孢子囊的致病力,以筛选出一种较好的保藏方法。结果表明,保藏6个月后,离体子叶冷冻法和离体叶片冷冻法保藏的孢子囊的萌发率、发病率和病情指数均较高;而10%二甲基亚砜+5%脱脂乳冷冻法保藏6个月后仅有2.16%的孢子囊能够萌发,菌株致病力丧失,发病率和病情指数均为0。保藏12个月后,离体子叶冷冻法保藏效果最好,孢子囊萌发率达到62.22%,发病率为90%,病情指数为48.89;离体叶片冷冻法保藏的孢子囊的致病力较弱;10%二甲基亚砜+5%脱脂乳冷冻法保藏12个月的孢子囊全部失活。此外,采用离体子叶冷冻法保藏12个月的32株寄生霜霉复壮成功率达到了93.75%。离体子叶冷冻法适用于寄生霜霉的保藏,其孢子囊可长时间保持较高的萌发率。     

小球藻的玻璃化超低温保存法

先用含0.5 m01·L-1甘油和0.4 mol·L-1蔗糖的预处理液处理20min,然后用含30%蔗糖 15%乙二醇 10%二甲基亚砜 BBMG培养液的玻璃化液处理,在0℃下预冻60min后,将小球藻投入液氮.此法存活率较高,可达到60.14%,小球藻种质保存效果较好.通过试验初步建立了小球藻玻璃化法超低温保存的技术程序.     

液氮保存曲霉菌种的效果

本文报告了以蒸馏水、10%甘油和5%二甲基亚砜(DMSO)为保护剂,用液氮冻结法保存5种8株曲霉的效果,并检测了这些菌分别产生的亚甲基丁二酸、柠檬酸、蛋白酶和糖化酶的生理活性。这些菌在液氮气相(接近-150℃)中保存180天全部保持着生活能力,它们的培养特征和形态特征保留原来的形状。所测定的液氮保存8株菌种的生理活性,除两株糖化酶活力稍有降低外,其它菌株没有明显的变化。     

外界因子对条斑紫菜自由壳孢子囊枝形成和生长影响

主要研究了不同光照周期(8L:16d、12L:12d和16L:8d)、不同温度(15℃、20℃和25℃)、不同光照强度(29、42、57、72和86μmol m-2·s-1)等外界因子对条斑紫菜自由丝状藻丝切断诱导形成壳孢子囊枝影响,以期获得条斑紫菜壳自由孢子囊枝形成的最佳外界条件.结果表明,在25℃和57μmol m-2·s-1条件培养30d后,短日照(8L:16d)对壳孢子囊枝诱导形成效果最好,其壳孢子囊枝形成率高迭92,5%,50d后几乎达到100%;在25℃和短日照条件培养30d后,光照强度为57μmol m-2·s-1,时,壳孢子囊枝诱导形成率达到最高(92.2%),光照强度过高过低均不利壳孢子囊枝形成;在57μmol m-2·s-1和短日照条件下,当温度为25℃时,壳孢子囊枝形成率最高,形成率随温度下降而减少.在12L:12d光照周期、57μmol m-2·s-1光照强度和25℃温度条件下,自由壳孢子囊枝生长最快,其日平均相对生长率可迭54.43%.以上研究为实现紫菜细胞工程育苗新技术一自由壳孢子囊枝育苗奠定了坚实基础.     

红花石蒜茎尖的玻璃化超低温保存

2～3mm的石蒜茎尖放在MS+0.4mol·L-1蔗糖的培养基上预培养5d,在25℃下用预处理液处理20min,接着用冰浴的玻璃化保护剂PVS2在冰浴中处理80min后,换新鲜PVS2并迅速投入液氮。液氮保存24h后,于40℃水浴中快速解冻2min,用MS+1.2mol·L-1蔗糖的液体培养基洗涤20min,滤纸吸干后接种到恢复培养基中,在25℃下暗培养7d后,转入光照强度为36μmol·m-2·s-1和光暗周期12/12h条件下培养。2周后的成活率最高可达90%,植株再生率达53%。     

SARS病毒NS-1株稳定性及培养条件的研究

用SARS病毒NS-1株接种Vero细胞,37℃培养,于培养1、2、3、4d收获病毒液,分别置-70℃冻融和4℃释放,于不同时期取样进行病毒滴定。结果显示,SARS病毒NS-1株各代次培养特性和形态学变化完全相同,有较好的抗原特异性,病毒滴度稳定,4℃保存125d,滴度下降2．25lgCCID50／ml,-70℃保存6个月,滴度未见明显下降。SARS病毒NS-1株未经冻融或释放,1d收获的病毒滴度比2、3、4d收获的病毒滴度低,经冻融或释放,1、2、3、4d收获的病毒滴度无明显区别,病毒收获时的病毒滴度与形态学变化成正相关。不同接种条件收获的病毒液,病毒滴度无明显区别。SARS NS-1株有较好的遗传稳定性和保存稳定性,培养2d,4℃释放收获病毒液,细胞悬液与病毒混合接种更简单,易操作,更适合疫苗规模生产。     

甘油包埋绿豆SOD的稳定性研究

用不同浓度的甘油包埋绿豆(Phczeolus rodiatus L.)超氧化物岐化酶(Superoxide dismutase,SOD),对不同条件下制备的甘油SOD的稳定性进行评估.结果表明:甘油包埋绿豆SOD的稳定性受甘油浓度影响明显,12.5%～25%为最适甘油浓度.在10℃、pH 6.8、搅拌速度1000 r min-1.条件下包埋的甘油SOD在55℃的平均半衰期为25.1 d,为非包埋SOD的5.1倍.甘油浓度为25%,温度分别为5℃、10℃、25℃、45℃条件下,pH 3.8中包埋的绿豆SOD耐酸性较好,pH 8.9下包埋的绿豆SOD的抗碱能力较强.在包埋过程中添加一定浓度的Zn2 、Cu2 、Fe2 有利于其热稳定性的提高.甘油包埋SOD对一些常见的化妆品添加剂也有一定的抗性.由此可见,甘油包埋技术可望作为一项有应用前景的SOD活性保持新技术,有利于极端温度和极端pH条件下的SOD的应用.     

大肠杆菌DH5α菌株感受态制备及转化率变化研究

通过氯化钙法制备大肠杆菌DH5α菌株感受态,讨论了不同保存温度和保存时间对感受态转化率的影响。结果表明,在4℃下保存,8h达到最高转化率;在-20℃和-70℃下保存,均为48h达到最高转化率。通过氯化钙法制备的DH5α菌株感受态细胞,在-20℃条件下简单保存,20d内完全可以满足一般转化研究的要求,不需要复杂的甘油、液氮处理及超低温要求。     

香蕉枯萎镰刀菌致病性快速测定方法的建立

建立致病性变异突变体库是研究香蕉枯萎镰刀菌致病机理的有效方法,此方法需要对大量突变体进行致病性测定,经典的根部接种测定方法耗时长达40 d,工作量十分巨大。因此,建立一种简便快速的致病性测定方法是高效建立香蕉枯萎病菌致病性变异突变体库的关键。本研究以香蕉巴西蕉和粉蕉叶片为材料,采用香蕉离体叶片分别接种香蕉枯萎病菌1号小种、4号小种、致病性丧失突变体、致病性严重减弱突变体,分别置于20℃、25℃、30℃、35℃、37℃条件下进行保湿培养3 d、5 d、7 d,观察发病情况,测定病斑大小。并通过根部接种法对离体叶片接种法的结果进行验证。结果表明,离体叶片测定法和根部接种法测定的结果一致,最佳致病温度30℃。从而建立了一种简便快速的香蕉枯萎病突变体致病性测定方法(离体叶片接种法)。运用该法在适宜的条件下可以准确、可靠、快速和简便测出香蕉枯萎病菌致病性,大大提高了突变体致病性测定筛选的效率。     

不同温度下白僵菌Bb04菌株对红火蚁工蚁的致病力

[背景[白僵菌是一种应用最广泛的虫生真菌,已被用于工厂化大量生产,可防治多种农林害虫。[方法]试验设置17、21、25、29和33℃5个温度水平,采用喷雾法,将浓度为1×105和1×108个·mL-1的白僵菌孢子悬浮液分别感染红火蚁工蚁,以测试不同温度下白僵菌菌株对红火蚁工蚁的致病力。[结果]当浓度为1×108个·mL-1时,在21、25和29℃条件下,红火蚁工蚁在15 d的累计死亡率均达到了100%,在17和33℃下分别为99.36%和98.74%。当浓度为1×105个·mL-1时,在21、25、29和33℃下,红火蚁工蚁在15 d的累计死亡率分别为29.42%、36.18%、33.17%和27.21%,显著高于17℃的累计死亡率。2个浓度处理在17～25℃时,白僵菌对红火蚁工蚁的致死中时（LT50）随着温度的升高而缩短,当温度为29和33℃时,红火蚁的LT50不减少反而增加。25℃时白僵菌Bb04菌株对红火蚁工蚁的LT50最短,死亡速度最快,致病力最强。[结论与意义]该研究可为制定田间利用白僵菌防治红火蚁的最佳时期奠定基础。     

Variation in Pathogenicity and Virulence of Strains of Xanthomonas campestris pv. glycines, the Incitant of Bacterial Pustule of Soybean

Standardized methods were developed to determine the pathogenicity and the degree of virulence of Xanthomonas campestris pv. glycines (Xcg) as well as the reaction of soybean plants in the greenhouse. A glass atomizer is described which allowed uniform inoculation without damaging the leaves. Optimum bacterial concentration was determined as 6 × 10⁶ CFU/ml for the pathogenicity test and 1.3 × 10⁵ CFU/ml for the virulence test. A total of 64 isolates were tested. Forty-five strains were designated as pathogenic, six of which were considered highly virulent. It was shown, for the first time, that large differences in the virulence of Xcg strains exist. All the highly virulent strains of Xcg were fresh isolates from diseased soybean leaves collected in Thailand. On the other hand, all the “old” cultures from bacterial collections possessed a low or very low virulence. Decrease of virulence of the pathogen did not occur very fast, however, that is: not within 2 years when stored on YDC-agar slants. Therefore, the bacteria may be kept on slants at 15 °C for short time storage, but the strains should be preserved permanently as lyophilized cultures.     

The Fate of Thiophanate-methyl Fungicide and Its Metabolites on Plant Leaves and Glass Plates

The persistence of thiophanate-methyl, dimethyl 4, 4′-o-phenylenebis (3-thioallophanate), and its metabolites on plant leaves and glass plates was examined using a radiolabeled compound. Apple and grape leaves and glass plates treated with thiophanate-methyl-phenyl-¹⁴C were kept outdoors, except on rainy days and nights when they were kept in green house.

Half lives of thiophanate-methyl on apple and grape leaves were around 15 and 12 days, respectively. On glass plates half life was around 3 days. The per cent of abundance of the parent compound and a major degraded product, methyl 2-benzirnidazolecarbamate, versus applied thiophanate-methyl at 14 days after treatment was as follows; 52.6 and 10.1% on apple leaves, 49.5 and 8.9% on grape leaves, and 5.5 and 24.1% on glass plates, respectively.     

枸杞木虱啮小蜂繁殖生物学研究

枸杞木虱啮小蜂Tamarixa lyciumiYang是枸杞木虱Paratrioza sinica Yang&Li若虫期的重要寄生性天敌。在实验室对其繁殖生物学特性进行了研究,结果如下:枸杞木虱啮小蜂大多进行两性生殖,孤雌生殖后代均为雄性,其自然性比为1.80:1。在15～35℃间,随温度升高,枸杞木虱啮小蜂发育历期缩短;从卵发育到成虫时需要8.22℃以上的有效积温217.21日·度。枸杞木虱啮小蜂在25℃恒温条件下繁殖力最高,35℃最低,寿命随温度的升高而缩短。在不同营养条件下,喂食20%蜂蜜的条件最适宜其繁殖,其次为20%蔗糖溶液,补充清水只可延长其寿命而不能提高繁殖力。5℃冷藏枸杞木虱啮小蜂蛹15d以内,不影响其正常羽化,冷藏20d或20d以上,羽化率显著降低;冷藏30d内对羽化后雌雄蜂寿命无明显影响,且从冷藏蛹中羽化的雌蜂寄生能力未受显著影响。     

Effects of kimchi extract and temperature on embryostasis of Ascaris suum eggs

To determine the effects of kimchi extracts at different temperatures on larval development, Ascaris suum eggs were mixed with soluble part of 7 different brands of commercially available kimchi and preserved at either 5℃ or 25℃ for up to 60 days. A. suum eggs incubated at 25℃ showed marked differences in larval development between kimchi extract and control group. While all eggs in the control group completed embryonation by day 21, only 30% of the eggs in the kimchi extract group became embryonated by day 36 and about 25% never became larvated even at day 60. At 5℃, however, none of the eggs showed larval development regardless of the incubation period or type of mixture group. To determine the survival rate of A. suum eggs that showed no embryonation after being preserved at 5℃, eggs preserved in kimchi extracts for 14, 28, and 60 at 5℃ were re-incubated at 25℃ for 3 weeks in distilled water. While all eggs in the control group became larvated, eggs in the kimchi extract group showed differences in their embryonation rates by the incubation period; 87.4 % and 41.7% of the eggs became embryonated after being refrigerated for 14 days and 28 days, respectively. When refrigerated for 60 days, however, no eggs mixed in kimchi extract showed larval development. Our results indicate that embryogenesis of A. suum eggs in kimchi extract was affected by duration of refrigeration, and that all eggs stopped larval development completely in kimchi kept at 5℃ for up to 60 days.     

Preliminary study on the cryopreservation of tropical bagrid catfish (Mystus nemurus) spermatozoa; the effect of extender and cryoprotectant on the motility after short-term storage

The effects of different extenders, and cryoprotectants on the motility of tropical bagrid catfish (Mystus nemurus) spermatozoa were evaluated after short-term storage. Three extenders, physiological saline, Ringer or saline at three levels of sperm to extender dilutions (1:20, 1:30, or 1:40) and four cryoprotectants (DMSO, ethanol, glycerol or methanol) at three concentrations (5, 10, or 15%) were examined in two separate experiments. In the first experiment, milt was suspended in the respective extender at the three milt to extender dilution ratios in two sets of tubes. Extended milt in the first set of tubes was stored at -4 degrees C, and motility assessed after 24h, while the second set was kept at 23 degrees C and sperm motility was assessed immediately and at 30-min intervals thereafter. Ringer retained sperm motility better than the other extenders at all dilution levels at temperatures of 23 and -4 degrees C respectively. At 23 degrees C, the sperm motility was almost completely lost after 150 min except for those in Ringer at 1:20 dilution level which still had a motility of 18% (compared to those kept at -4 degrees C for 24, which had motility from 39 to 71%, regardless of extender). In the second experiment, various cryoprotectants were added to solutions of milt (that was diluted in Ringer at 1:20 ratio and cryopreserved in liquid nitrogen for 15 days). Sperm cryopreserved in 10% methanol had the highest motility (58%) compared with those in the other cryoprotectants at all concentrations.     

人工扩繁代异色瓢虫卵和成虫最适冷藏条件的探讨

探讨了人工扩繁异色瓢虫时卵和成虫的最适冷藏条件。卵的最适保存温度是 1 0℃ ,冷藏 1 5d内孵化率平均在 60 %以上。成虫的最适保存温度是 1 0℃ ,如将刚羽化的成虫直接在该温度下冷藏 ,冷藏 40d时存活率能保持在 5 0 %左右 ;如将刚羽化成虫先在 1 5℃、0L∶2 4D条件下处理 1 6d,然后置于 1 0℃下冷藏 ,经冷藏 70d后存活率接近 1 0 0 % ,冷藏 90d后存活率仍高于 70 %。表明经一定条件下预处理后再冷藏成虫能保持较高的存活率和较长的存活期 ,可满足人工扩繁时对成虫的冷藏要求。     

The development of Puccinia hordei on barley cv. Zephyr

Germination of uredospores of Puccinia hordei was similar on cover-slips and on the first leaves of barley seedlings (cv. Zephyr) at 100 % r.h. over the range 5–25 °C, being greatest at 20 °C. At 15, 20 and 25 °C maximum germination was attained in 6 h. No uredospores germinated on coverslips in humidities below saturation. The numbers of pustules which subsequently developed on plants incubated at 5, 10, 15 or 18 °C and 100 % r.h. for varying periods up to 24 h, were directly related to rise in temperature and length of incubation. The time from inoculation to eruption of pustules (generation time) was 6 days at 25 °C, 8 days at 20 °C, 10 days at 15 °C, 15 days at 10 °C and 60 days at 5 °C. Pustule production on inoculated plants which had been kept at 5 °C was rapidly accelerated when they were transferred to 20 °C. Data obtained at constant temperatures were used to predict generation times of the fungus in the field. The productivity of pustules, determined as weight of uredospores, was examined at 10, 15 and 20 °C. Significantly more spores were produced at 15 than at 10 °C and most were produced at 20 °C. The results are discussed in relation to those obtained by other workers and to the development of brown rust in the field.     

The biology of Peronospora viciae on pea: laboratory experiments on the effects of temperature, relative humidity and light on the production, germination and infectivity of sporangia

Germination of Peronospora viciae sporangia washed off infected leaves varied from 20% to 60%. Sporangia shaken off in the dry state gave 11–19% germination. Most sporangia lost viability within 3 days after being shed, though a few survived at least 5 days. Infected leaves could produce sporangia up to 6 weeks after infection, and sporulating lesions carried viable sporangia for 3 weeks. Sporangia germinated over the range 1–24 °C, with an optimum between 4 and 8 °C. Light and no effct. The temperature limits for infection were the same as for germination, but with an optimum between 12 and 20 °C. A minimum leaf-wetness period of 4h was required, and was independent of temperature over the range 4–24 °C. Maximum infectivity occurred after 6h leaf wetness at temperatures between 8 and 20 °C. Infection occurred equally in continuous light or in darkness. After an incubation period of 6–10 days sporangia were produced on infected leaves at temperatures between 4 and 24 °C, with an optimum of 12–20 °C. Exposure to temperatures of 20–24 °C for 10 days reduced subsequent sporulation. Sporangia produced at suboptimal temperatures were larger, and at 20 °C. smaller, than those produce at 12–16 °C. Viability was also reduced. No sporangia were produced in continuous light, or at relative humidities below 91%. For maximum sporulaiton an r.h. of 100% was required, following a lower r.h. during incubation. Oospores wre commonly formed in sporulating lesions, and also where conditons limited or prevented sporulation. The results are discussed briefly in relaiton to disease development under field conditions.